Methods and devices for analysis of sealed containers

ABSTRACT

This invention provides methods, NMR probes, and NMR systems for the analysis of the contents of sealed food and beverage containers and the like.

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to and benefit of U.S.applications Ser. No. 60/396,644, filed Jul. 17, 2002 and 60/465,644,filed Apr. 25, 2003, the full disclosures of which are incorporatedherein by reference.

FIELD OF THE INVENTION

[0002] This invention relates to methods and devices for analyzingsealed food and beverage containers, and particularly sealed winebottles, by NMR spectroscopy.

BACKGROUND OF THE INVENTION

[0003] Wine is the product of the growth and metabolism of yeasts andbacteria in grape must. It is well known that many of these and otherbacteria survive all of the steps of wine making from the mature grapethrough vinification to bottle corking (Ribereau-Gayon (1985) “Newdevelopments in wine microbiology” Am. J. Enol. Vitic. 36:1-10). Oneclass of organisms of interest is Acetobacter, a bacteria responsiblefor oxidizing ethyl alcohol into vinegar or acetic acid (Fleet andDrysdale (1988) “Acetic acid bacteria in winemaking: a review” Am. J.Enol. Vitic. 39:143-154; Lonvaud-Funel and Millet (2000) “The viable butnon-culturable state of wine micro-organisms during storage” Lt. Appl.Microbiol. 30:136-141). Although present in most wines, Acetobacter doesnot typically generate enough acetic acid to spoil wine during bottlestorage due to a lack of oxygen. As long as the wine is stored in ananaerobic environment, conditions ensured by quality corking, acceptablylow quantities of acetic acid (e.g., below sensory levels) are producedand the quality of the wine is preserved. Unfortunately, the sealingperformance of wine corks can degrade with age, and the long termbehavior of low quality natural corks and synthetic stoppers is not welldocumented. One consequence of a leaky cork is the admission of oxygento wine, a triggering of Acetobacter function, and the production ofacetic acid . Furthermore, the admission of oxygen into the bottle inthe presence of heat can lead to oxidation of ethanol into aldehydes.These processes lead to changes in odor and flavor, and thereforespoilage, of fine wines.

[0004] Current methods for identifying acetic acid in wine are verysensitive, detecting roughly 50 μg/L acetic acid, even though theaccepted spoilage limit of acetic acid in wine is 1.4 g/L (see, forexample, Kellner et al. (1998) “High performance liquid chromatographywith real-time Fourier-transform infrared detection for thedetermination of carbohydrates, alcohols and organic acids in wines” J.Chromatogr. A. 824:159-167; Garcia-Martinet al. (2000) “Simultaneousdetermination of organic acids in wine samples by capillaryelectrophoresis and UV detection: optimization with five differentbackground electrolytes” J. High Resol. Chromatogr. 23:647-652; Kellneret al. (1998) “A rapid automated method for wine analysis based uponsequential injection (SI)-FTIR spectroscopy” Fresenius 362:130-136; andMargalith (1981) in Flavour Microbiology, pp. 167-168, Charles ThomasPublishers, Springfield, Ill.). In addition, nuclear magnetic resonance(NMR) spectroscopy has been employed for wine fingerprinting studies andtrace amino acid and organic molecule detection in wine (Guillou andReniero (1998) “Magnetic resonance sniffs out bad wine” Physics World11:22-23; and Kidric et al. (1998) “Wine analysis by 1D and 2D NMRspectroscopy” Analysis 26:97-101). However, all published NMR studies ofwine involve removal and analysis of small volume samples of wine (e.g.,less then 1 mL) to accomplish these measurements. As such, all of thecurrent strategies for contaminant (e.g., acetic acid) detection requirethe bottle to be violated, a process that destroys the cork, seal, andlabel, severely devaluing both the wine and bottle. The presentinvention overcomes these and other problems by providing methods anddevices for the detection of contaminants in wine bottles by NMRspectroscopy. These methods are equally applicable to other sealedconsumables containers for which contamination, degradation, or otherchanges in product flavor or quality is a concern.

SUMMARY OF THE INVENTION

[0005] The present invention provides methods and devices for theanalysis of sealed consumable containers by NMR spectroscopy. The highstatic and radiofrequency (rf) magnetic fields used in the NMRexperiment in no way affect the quality of the food or beverage examinedvia the methods provided herein.

[0006] In some embodiments, the present invention provides non-invasive,non-destructive analytical methods for determining the level of wineacetification. As such, the methods and devices of the present inventioncan be routinely used in the evaluation of the quality of fine wines andin the study of wine cork aging. Furthermore, these methods of intactbottle analysis are not limited to the determination of acetic acidspoilage and content in wines, but can be extended to the study of otherwine molecular components (e.g., aldehydes and flavenoids), as well asto components and/or contaminants in other types of sealed consumables.

[0007] Accordingly, the present invention provides methods for analyzingone or more contents of a sealed consumables container. The methodsinclude, but are not limited to, the steps of providing an NMRspectrometer and an NMR probe configured to accept a portion of thesealed consumables container; positioning the portion of the sealedconsumables container within a data collection region of the NMR probe;establishing a homogeneous static magnetic field across the datacollection region; collecting an NMR spectrum; and analyzing one or morepeaks in the NMR spectrum, thereby analyzing one or more contents of thesealed consumables container.

[0008] Any food or beverage having components that generate one or moreNMR peaks can be assessed using the methods and devices of the presentinvention. Thus, a variety of food or beverage containers having, forexample, nonalcoholic beverages, alcoholic beverages, beer, vinegar orolive oil stored therein, can be analyzed using the methods of thepresent invention. In a preferred embodiment, the sealed consumablescontainer is a bottle of wine.

[0009] The methods of the present invention can be used in a qualitativeor quantitative manner, e.g., either the presence of a selectedcomponent or the concentration of the selected component is determined.For example, in the analysis of wine, exemplary selected componentsinclude, but are not limited to, acetic acid, aldehydes, flavenoids, andamino acids.

[0010] The methods of the present invention include the step ofpositioning the portion of the consumables container within a datacollection region of the NMR probe. For example, either the neck of thecontainer or a portion of the body of the container can be placed withinthe data collection region of the NMR probe. The homogeneous staticmagnetic field is then established across the data collection region by,for example, adjusting the one or more shim coils in the probe.Preferably, establishing the homogeneous field allows for resolution ofchemical shift difference between selected NMR spectra peaks a minimumdistance apart. In certain embodiments of the present inventioninvolving ¹H NMR spectroscopy, the resolution will preferably allow fordistinguishing peaks that are about 1 ppm apart. Optionally, the NMRpeaks generated by the selected components are integrated, therebyanalyzing a quantity of the selected component.

[0011] The present invention also provides NMR probes configured toposition a portion of a sealed consumables container within an NMRspectrometer. The NMR probes used in the present invention can be any ofa number of detection probes, including, but not limited to, a ¹H probe,a ²H probe, a ¹³C probe, a ¹⁷O probe, or a combination thereof. The NMRprobe components include a body structure having a cavity adapted forreceiving a portion of the sealed consumables container (e.g., a neck ofa bottle, or a body of the container). The cavity is typically disposedin the body structure (either at a first end, or in a middle portion),such that a first rf coil attached to the body structure is positionedproximal to the cavity and the portion of the sealed container. In oneembodiment of the probes of the present invention, the first rf coilcomprises a split solenoid coil, in which the coil portions arepositioned to either side of the data collection region of the probe. Inan alternate embodiment, the first rf coil is a birdcage-style coilsurrounding the data collection region of the probe.

[0012] In some embodiments of the present invention, the first rf coilis used for both transmitting and receiving rf pulses. Optionally, theprobe includes a second rf coil positioned distal to the first rf coil.The second rf coils can be, for example, configured for measurement ofone or more signals from a calibration sample. Alternatively, the secondrf coil is configured for selective excitation of a heteronucleus, suchas ¹³C, ¹⁷O, ²H, ²³Na, ²⁷Al, ¹⁹⁹Hg, or ²⁰⁷Pb.

[0013] The probes of the present invention fuirther include a tuningcapacitor coupled at a first position to the rf coil, and coupled at asecond position to a length of coaxial cable configured for connectionto the NMR spectrometer. The tuning capacitor can include, but is notlimited to, one or more non-magnetic zero-to-ten (0-10) picofarad highpower rf capacitors.

[0014] Optionally, the probe also includes additional components usefulfor NMR analyses, such as electronic components for generating magneticfield gradients, a calibration fluid sample tube; and a fluid jacket formodulating the probe temperature, to name a few.

[0015] Systems for analyzing contents of a sealed consumables containerare also provided by the present invention. The system componentsinclude, but are not limited to, the NMR probe configured to position aportion of a sealed consumables container within an NMR spectrometer; anNMR spectrometer having a bore proximal to a magnet and configured toreceive the NMR probe, an amplifier coupled to the NMR probe viaco-axial cable; and a receiver system having a preamplifier and adetector. Optionally, the system further includes a pulse programmer.

[0016] Optionally, the NMR probe of the system is a single resonanceprobe selected from the group consisting of a ¹H probe, a ²H probe, a¹³C probe, an ¹⁷O probe, a ²³Na probe, an ²⁷Al probe, a ¹⁹⁹Hg probe, anda ²⁰⁷Pb probe. In one embodiment, the NMR probe employs a first rf coilused for both transmitting and receiving rf pulses. In another aspect,the NMR probe further comprises a second rf coil configured, forexample, for measurement of one or more signals from a calibrationsample.

[0017] The NMR probe is configured to accept the sealed consumablescontainer and position a portion of the container (e.g., the neck of abottle, or the body of the container) within the magnetic field of thespectrometer. Typically, the spectrometer comprises a wide bore magnet;preferably, the magnetic field is generated by a room temperaturesuperconducting magnet. While any field strength can be used in thesystem of the present invention, higher field strengths are preferableto lower field strengths. In one embodiment, magnetic field comprises a2.01 T magnetic field. The receiver component of the analytical systemincludes, but is not limited to, preamplifier and a detector incommunication with the NMR probe. In one embodiment, the receiverincludes a passive rf duplexer and signal mixing and digitizationelectronics. These and other aspects of the present invention areprovided herein.

BRIEF DESCRIPTION OF THE DRAWINGS

[0018]FIG. 1 is a schematic drawing of an exemplary probe of the presentinvention.

[0019]FIG. 2 depicts an expanded view of an exemplary probe, showing theplacement of a sealed container within the data collection region.

[0020]FIG. 3 panels A and B represent one embodiment of the systems ofthe present invention, depicting an experimental setup used to obtain anNMR spectrum of a full, intact wine bottle. FIG. 3A provides a schematicdepicting the placement of the sealed consumables container (a winebottle) and NMR probe within the body structure of an NMR spectrometer.FIG. 3B shows an expanded view of the probe, depicting the positioningof the selected portion of the container with the rf coils of the probe,and indicating that the NMR probe head is capable of housing an entirebottle of wine.

[0021]FIG. 4 panels A and B depict an alternative embodiment of thesystems of the present invention, showing the placement of the body ofthe sealed consumables container within the NMR probe. FIG. 4B shows anexpanded view of the probe, depicting the positioning of the body of thecontainer within the sample measurement region of the probe.

[0022]FIG. 5 depicts NMR spectral data obtained at 9.1 T for a 500 μLsample of wine (panel A) and red wine vinegar (panel B).

[0023]FIG. 6, panels A and B, provides spectral data generated for asample of wine (panel A) and a sample of red wine vinegar (panel B)using the methods and probes of the present invention.

[0024]FIG. 7 provides a plot comparing the experimental versuscalculated values of acetic acid provided in a set of acetic acid/ethylalcohol full bottle standard samples.

[0025]FIG. 8 panels A, B and C provide exemplary rf pulse sequences foruse in the methods of the present invention.

[0026]FIG. 9 panels A and B depict ¹³C NMR spectra on full bottles ofwine.

[0027]FIG. 10 panels A and B are tables depicting NMR-derivedpercentages of ethanol (FIG. 10A) and acetic acid concentrations (FIG.10B) in a vertical series of sealed full bottles of the UC DavisCabernet Sauvignon bottled between 1950 and 1977.

DETAILED DESCRIPTION

[0028] DEFINITIONS

[0029] Before describing the present invention in detail, it is to beunderstood that this invention is not limited to particular devices orcontainer systems, which can, of course, vary. It is also to beunderstood that the terminology used herein is for the purpose ofdescribing particular embodiments only, and is not intended to belimiting. As used in this specification and the appended claims, thesingular forms “a”, “an” and “the” include plural referents unless thecontent clearly dictates otherwise. Thus, for example, reference to “acapacitor” includes a combination of two or more capacitors; referenceto a “coil” includes mixtures or series of coils, and the like.

[0030] Unless defined otherwise, all technical and scientific terms usedherein have the same meaning as commonly understood by one of ordinaryskill in the art to which the invention pertains. Although any methodsand materials similar or equivalent to those described herein can beused in the practice for testing of the present invention, the preferredmaterials and methods are described herein. In describing and claimingthe present invention, the following terminology will be used inaccordance with the definitions set out below.

[0031] The term “consumables” as used herein refers to a food, beverage,or alternate energy source (e.g., bacterial media) intended forconsumption by an organism (e.g., a human, an animal, a cell culture,and the like). Thus, the term “sealed consumables container” refers to apackaged or unopened vessel or receptacle containing the food orbeverage. Sealed NMR tubes prepared with samples of food or beverageproducts are not considered sealed consumables containers with respectto the present invention.

[0032] The terms “NMR probe” and “probe head” are used interchangeablyherein to refer to the component of an NMR spectrometer system whichtransmits pulses to the sample and receives the NMR signals generated.

[0033] The term “data collection region” refers to the portion of theNMR probe in which the NMR signal is generated; typically, thehomogeneity of the magnetic field of the spectrometer is optimized inthis region.

[0034] The term “rf coil” refers to a set of filamentary wire sectionsarranged in a helical geometry and designed for transmitting and/orreceiving radiofrequency signals.

[0035] The term “tuning capacitor” as used herein refers to one or morecapacitor components of the NMR probe which are typically used to matchand tune the probe to the correct Larmor frequency and impedance matchthe rf circuit to, for example, 50 Ω.

[0036] The term “split solenoid coil” (or “split pair solenoid”) refersto a solenoid having multiple coils of wire (usually in cylindricalform) that generates a magnetic field when carrying a current.

[0037] METHODS

[0038] The present invention provides methods of analyzing one or morecontents of a sealed consumables container. The sealed consumablescontainer can be any of a number of food or beverage containers havingcontents of interest, including, but not limited to, alcoholic ornonalcoholic beverages.

[0039] In a preferred embodiment, the container is a corked (e.g.,unopened) bottle of wine. Any number of wine bottle “styles” can beaccommodated in the methods (as well as devices and systems) of thepresent invention. For example, the methods of the present invention canbe used to analyzed the contents of the high shouldered “Bordeaux”bottle (typically used for Sauvignon Blanc, Cabernet Sauvignon, Merlot,and Bordeaux blends), the slope shouldered “Burgundy” bottle (Chardonnayand Pinot Noir), or the taller “Hoch” bottle of Germanic origin(Rieslings and Gewiirztraminers). The contents of champagne/sparklingwine bottles, Chianti bottles, and the shaped-neck bottles typicallyused for fortified wines (port, sherry, etc.) can also be analyzed bythe methods of the present invention. Furthermore, a range of bottlesizes can be used in the methods of the present invention; in additionto the 750 mL bottle found in typical wine cellars, the smaller halfbottles, “splits” (187 mL) and “tenths” (375 mL) as well as the larger“magnum” bottles (e.g., 1L, 1.5L and 3L bottles) can be examined.

[0040] The bottle can be made of either clear or colored (e.g., amber,green or brown) glass. In addition, the beeswax seal and/or lead capoften used in the corking process need not be removed for the analysis.

[0041] In addition to wine, other consumables can be analyzed by themethods of the present invention, including, but not limited to, beer,vinegar and olive oil. In addition, sealed receptacles containingsolutions or suspensions not typically considered as “food” (e.g.,microbial culture media, herbal tinctures, and the like) can also beexamined using the methods of the present invention. Preferably, thecomponent of interest in the sealed container generates an NMR spectrumhaving at one or more sharply defined peaks.

[0042] The methods of analyzing one or more contents of the sealedconsumables container employ an NMR spectrometer and an NMR probeconfigured to accept a portion of the sealed consumables container. Inone embodiment of the present invention, the NMR probe is configured toreceive the narrowed upper portion, or “neck,” of the sealed container.In an alternate embodiment, the body of the container is the portionplaced in the NMR probe.

[0043] In the methods of the present invention, the selected portion ofthe sealed consumables container is positioned within the datacollection region of the NMR probe. This can be achieve by placing thecontainer within the probe, and then inserting the probe into thespectrometer, such that the selected portion of the container (neck,body, etc.) is optimally positioned within the magnetic field of thespectrometer. Alternatively, the probe can be installed into thespectrometer prior to insertion of the container. In either case, thecontainer is positioned such that a portion of the consumables ispositioned within the magnetic field of the spectrometer, and proximalto the rf coil of the NMR probe. The examined portion of the sealedcontainer will be determined in part by the shape and/or configurationof the sealed container, as well as various requirements with respect tothe type of NMR spectroscopy performed. For example, either the neck ofthe container or a portion of the body of the container can be placedwithin the data collection region of the NMR probe.

[0044] In one preferred embodiment, the rf coil “examines” the neck of awine bottle between the base of the cork and the flared body of the winebottle. Although there is less sample in this region (and therefore lesssignal) as compared to the larger body of the bottle, it is easier toestablish a homogeneous static magnetic field over this smaller sampleregion, thus enhancing the probability of obtaining narrow (resolved)NMR spectral peaks.

[0045] A homogeneous static magnetic field is established across thedata collection region of the NMR probe by standard mechanisms, e.g., byadjustment of cryogenic and/or room temperature (RT) magnetic fieldshims. The NMR spectrum is then collected by monitoring the response ofthe sample to an rf electromagnetic field pulse generated by the rfcoil. Preferably, the magnetic field established is homogeneous enoughto allow for resolution of chemical shift differences between selectedNMR spectra peaks set a minimum distance apart. The degree ofhomogeneity necessary for performing the methods of the presentinvention will depend on a number of factors, including nucleiselection, magnetic field strength, and chemical structure. In themethods of the present invention, the homogeneous static magnetic fieldis established such that one or more peaks of interest from thecontaminant are resolved from additional NMR spectral peaks. Forexample, for ¹H NMR spectra collected on the contents of sealed winebottles, the minimum desired resolution is approximately 1 ppm, thedistance between the methyl resonance and the methylene resonance of theacetic acid contaminant. Exemplary NMR spectra of a number of compoundscan be found, for example, in the Aldrich Library of ¹ H and ¹³ C FT NMRSpectra, Edition I (1993, volumes 1-3, eds. Pouchert and Behnke, AldrichChemical Company), from which a desired minimum resolution can bereadily determined by one of skill in the art.

[0046] Since the magnetic field is not stabilized with a flux-lockedloop, and a ²H lock (as typically employed with small volume NMR samples“spiked” with a deuterated standard such as TMS) is not possible forsealed wine bottles, data collection is typically performed via blockaveraging (e.g., n data sets of free induction decay each derived from mscans). In a preferred embodiment, the data are collected as blockaverages of n=10 groups of 100 scans. The n=10 free induction signalsare Fourier transformed, overlapped by shifting the frequency, and addedoffline using Matlab (Mathworks Inc, Natick Mass.). This procedureeliminates the effect of the long time drift in the static magneticfield on the collected data, thereby producing highly resolved ¹H NMRspectra for the methyl group region in wine.

[0047] After an NMR spectrum is collected, the peaks of the spectrum areexamined. Typically, the analysis involves examination ofpreviously-identified peaks in a select region of the spectrum. Thepeaks can represent any of a number of components found in the sealedcontainer. For example, the peaks of interest are optionally generatedby contaminating molecular species (contaminants) indicating spoilage orexposure to oxygen. For embodiments involving the analysis of wine, oneparticular contaminant of interest is acetic acid, which is generated bythe bacterial metabolism of ethyl alcohol. For analysis of acetic acid,the regions of interest are around 1 ppm (the region in which the methylpeak for acetic acid can be found) as well as around 3.6 ppm (the regionin which the methylene peak from acetic acid is located). Alternatively,wine components such as aldehydes or flavenoids can be examined.

[0048] In some embodiments of the method, the analysis is on aqualitative level, e.g., are the NMR peaks of interest present orabsent. In other embodiments, the analysis is quantitative; the selectedpeaks are integrated and compared to a standard peak intensity, therebyproviding a quantitative analysis of the selected components of thesealed consumables container. Preferably, the NMR resonances generatedby the component of interest are sharp, facilitating the optionalintegration process. The integration can be performed using a softwareprogram provided with the spectrometer operational software, or it canbe performed the old-fashioned way, by printing the spectra, cutting outthe peaks of interest, and weighing the paper scraps.

[0049] NMR PROBES

[0050] The present invention also provides NMR probes for use in themethods described herein. The NMR probes of the present invention areconfigured to position a portion of a sealed consumables containerwithin an NMR spectrometer, thus avoiding the need to violate the sealon the container in order to analyze the contents. The probes typicallycomprise a body structure having a cavity disposed at a first end of thebody structure, a first rf coil positioned proximal to the cavity andthe portion of the sealed container; and a tuning capacitor coupled tothe rf coil and to a length of coaxial cable configured for connectionto the NMR spectrometer. In an alternate embodiment, the cavity isdisposed in a middle region of the body structure, rather than proximalto the end of the probe.

[0051] The probes of the present invention can be used to detect anydesired nuclei capable of generating a nuclear magnetic resonance andhaving adequate chemical shift dispersion between selected contaminantand/or sample signals. Thus, the probes of the present inventioninclude, but are not limited to, ¹H probes, ²H probes, ¹³C probes, ¹⁷Oprobes, and the like. Furthermore, the probes of the present inventioncan be single frequency or dual frequency probes (e.g., a ¹H/¹³C probe).

[0052] The body of the probe is typically composed of material having alow magnetic susceptibility to reduce and/or prevent distortion of thestatic magnetic field when the probe is positioned in the spectrometer.Exemplary materials used in the manufacture of the body structure (orportions thereof) include, but are not limited to stainless steel,aluminum, glass, ceramic, and plastics such as Teflon(polytetrafluoroethene), Kel-F (polychlorotrifluoroetene), and PVC(polyvinylchloride).

[0053] The body structure has a cavity that is configured to accept aportion of the sealed consumables container, such that a portion of thecontainer is positioned within the data collection region of the probe.Thus, the sample cavity is greater than that typically employed in anprobe configured for NMR tubes. The overall dimensions of the probeoptionally range from about 600 mm to 800 mm in length, preferably about700 mm. The outer diameter of the probe ranges in size from 100 mm to150 mm in diameter, although an outer diameter of up to 310 mm ispossible with the current magnet embodiment. The size of the cavityportion of the probe will depend upon the sealed container to beanalyzed; for a probe configured to accept a neck portion of a winebottle (FIGS. 3A and 3B), the cavity portion of the probe will typicallyrange from 34 mm to 85 mm in diameter. Larger cavities able to encompassa wider portion of a consumables container, such as the base and body ofa wine bottle (e.g., about 100-150 mm in diameter), are alsocontemplated (see FIGS. 4A and 4B).

[0054] The cavity is configured to hold the sealed container in positionthrough the use of, for example, one or more PVC positioning rings. Inone embodiment, the cavity extends from one end of the probe to the datacollection region, for insertion of the sealed container from the openend. In an alternate embodiment, the cavity is enclosed within the bodystructure, and accessed by an opening in the side of the body structure.

[0055] The first rf coil is positioned in the body structure of theprobe, proximal to the cavity (and the selected region of the sealedcontainer inserted therein). Optionally, the first rf coil functions asboth the transmitting coil and the receiving coil. In one embodiment,the first rf coil is a split solenoid coil. An exemplary split solenoidcoil is 12 gauge copper wire wound in a 1 cm diameter spiral, the firstcoil portion having 4 turns of the copper wire, and coupled (via aconnecting portion of the wire) to a second coil portion having another4 turns of copper wire. The first coil portion is positioned on one sideof the cavity, while the second coil portion is positioned on anopposite side of the cavity; the connecting wire runs between the twoportions without crossing the cavity itself (e.g., along thecircumference of the cavity). Preferably, the second coil portioned isaligned along a same axis as the first coil portion.

[0056] In another embodiment, the rf coil circumscribes the cavity(e.g., the walls of the body structure defining the cavity act as aformer around which the rf coil is wound.) In a further embodiment ofthe probe, the first rf coil comprises a birdcage-style coil. Such aconfiguration of coil portions is described in, for example, Hayes etal. (1985) “An efficient, highly homogeneous radiofrequency coil forwhole-body NMR imaging at 1.5 T” J. Magn. Reson. 63:622-628.

[0057] The probes of the present invention also include one or moretuning capacitors. The tuning capacitor is coupled at a first positionto the first rf coil, and coupled at a second position to a length ofcoaxial cable configured for connection to the NMR spectrometer. In oneembodiment, the tuning capacitor is a non-magnetic 0-10 picofarad highpower rf capacitor.

[0058] A schematic representation of the probes of the present inventionis shown in FIG. 1. Probe 10 comprises body structure 20, first rf(radiofrequency) coil 30; and tuning capacitors 40 and 42. Bodystructure 20 has opening or cavity 50 disposed at one end for receivingthe sealed consumables container (not shown).

[0059] A portion of cavity 50 extends into data collection region 60 ofprobe 10. First rf coil 30 is attached to capacitor 40 at a first end 32and attached to capacitor 42 at a second end 34, and is positionedproximal to cavity 50 such that coil portions 36 and 38 are situated toeither side of data collection region 60.

[0060] Tuning capacitors 40 and 42 are also coupled at second positions44 and 46 to coaxial cables 70 and 72, which are configured forconnection to the NMR spectrometer (not shown). In addition, tuningcapacitor 42 is coupled at a third position to rf in/out coaxial cable74, which provides the radiofrequency signal for NMR spectrumgeneration.

[0061]FIG. 2 depicts an expanded view of exemplary probe 110, showingthe placement of sealed container 100 within the data collection region160. Coil portions 136 and 138 of rf coil 130 are approximately 2.0 cmin diameter (measurement A) and extend approximately 2.5 cm from theupper surface of tuning capacitors 140 and 142, respectively(measurement B), such that the total height of rf coil 130 isapproximately 4.5 cm. Coil portions 136 and 138 are positionedapproximately 3.4 cm apart (measurement C) with the intermediate coilportion (represented by dotted line) arcing between the two portions,such that neck portion 102 of sealed container 100 can be positionedbetween coil portions 136 and 138 for optimal data collection.Optionally, container 100 will have stopper 104 positioned at the distalend of neck portion 102. Stopper 104 is optionally a cork, a screw-topcap, or a plug. Preferably, bottle 100 is positioned within datacollection region 160 such that stopper 104 does not interfere with thedata collection procedure.

[0062] Probe 110 optionally includes positioning ring 112 separating rfcoil 130 from the main portion of cavity 150; the aperture inpositioning ring 112 allows the selected portion of bottle 100 to bepositioned within data collection region 160 while protecting thisregion from dust, etc. Optional capacitor stand 114 is positioned on thedistal side of tuning capacitors 140 and 142. Capacitors 140 and 142 areapproximately 4.5 cm in height; therefore, the distance between a faredge of coil portion 136 and the distal side of capacitor 140 isapproximately 9 cm, and the distance between outer edges of positioningring 112 and capacitor stand 114 is approximately 11 cm.

[0063] The probes of the present invention optionally incorporate asecond rf coil, preferably positioned distal to the first rf coil. Thesecond rf coil can be employed for a number of purposes. For example,the second rf coil can be used for either transmitting or receiving therf signal (in embodiments in which the first rf coil does not functionas both transmitter and receiver). Alternatively, the second rf coil canbe configured for measurement of one or more signals from a calibrationsample. In yet another embodiment, the second rf coil provides forselective excitation of a heteronucleus (including, but not limited to,¹³C, ¹⁷O, ²H, ²³Na, ²⁷Al, ¹⁹⁹Hg, ²⁰⁷Pb, and the like).

[0064] Optionally, the probe further includes one or more components fortuning and/or impedance matching the rf coil(s) to at least one rf powersource at a selected frequency.

[0065] The probes of the present invention optionally include one ormore additional components which enhance the functioning of the probe.For example, the probe can include components for generating magneticfield gradients, which can be used, for example, for imaging purposes.In some embodiments, the probe includes a calibration fluid sample tube.The optional calibration sample tube is typically positioned within thecavity of the body structure such that the calibration sample ispositioned proximal to the selected portion of the sealed consumablescontainer when the container is inserted in the cavity.

[0066] In a further embodiment, the NMR probes of the present inventionoptionally further include a fluid jacket, reservoir or other mechanismfor modulating the temperature of the probe. Exemplary fluid jacketdesigns for use with the present invention are described in, forexample, U.S. Pat. No. 5,530,353 titled “Variable Temperature NMR Probe”(Blanz).

[0067] SYSTEM COMPONENTS

[0068] The present invention also provides systems for analyzingcontents of a sealed consumables container. The systems include one ormore NMR probes of the present invention, an NMR spectrometer, and areceiver system configured for electronic communication with the NMRprobe. The probes and systems of the present invention can be used toperform pulsed, continuous wave, or gradient NMR experiments.

[0069] The NMR spectrometer typically comprises a body structure, amagnet housed within the body structure, a bore proximal to the magnetand configured to receive the NMR probe, and an amplifier configured forcoupling to a first position on the NMR probe. Optionally, the magnet isa constant external magnet, a room temperature (RT) magnet, and/or asuperconducting magnet. Any NMR spectrometer having a bore capable ofreceiving the NMR probes can be used in the systems of the presentinvention. Preferably, the NMR spectrometer is a super wide borespectrometer. Exemplary spectrometers are available commercially from,for example, Varian (Palo Alto, Calif.; www.varianinc.com) and Bruker(Germany, www.bruker.com). The field strength of the magnet componentused in the systems can also vary, ranging from 2.01 T to 9.4 T andhigher.

[0070] The systems of the present invention include a receiver systemconfigured for electronic communication with the NMR probe. Optionally,the receiver system is incorporated into the body structure of the NMRspectrometer. The receiver system typically comprises a preamplifierconfigured for coupling to the NMR probe and a detector in communicationwith the preamplifier. In one embodiment of the systems of the presentinvention, the receiver includes a passive rf duplexer as well aselectronics for signal mixing and digitization (see, for example,Fukushima and Roeder, Experimental Pulse NMR a Nuts and Bolts Approach,Addison-Wesley, New York, 1981).

[0071] Optionally, the system further includes an NMR pulse programmer.Exemplary pulse programmers are available from Tecmag, Inc. (Houston,Tex.; www.tecmag.com).

[0072] In some embodiments of the present invention, the system includesa mechanism for spinning the sealed container within the NMR probe.Exemplary spinning mechanisms include, but are not limited toair-propelled mechanisms (e.g., air turbines), rotor mechanisms,strap-based mechanisms and the like.

[0073] In a preferred embodiment of the present invention, the systemalso includes a rf power source, for exciting the nuclei within thesealed container.

[0074]FIG. 3A provides an exemplary system of the present inventiondepicting the positioning of bottle 200 within the data collectionregion 260 of probe 210, which is inserted into magnet 280 of the NMRspectrometer. FIG. 3B shows the alignment of bottle 200 within probe 210with respect to rf coil 230 and tuning capacitors 240 and 242. Alsodepicted are optional components positioning ring 212 and capacitorstand 214.

[0075]FIGS. 4A and 4B depict an alternate positioning of bottle 300within the data collection region of probe 310, in which the body ofbottle 300 is inserted into data collection region 360. In FIG. 4A,probe 310 is inserted into magnet 380 of the NMR spectrometer. FIG. 4Bshows rf coil 330, tuning capacitors 340 and 342, coaxial cables 370 and372, and rf in/out cable 374, with respect to the alignment of bottle300 within probe 310. Positioning ring 376 centers the sample inside ofrf coil 330, which is mounted on PVC positioning rings 378 and 379.

EXAMPLES

[0076] It is understood that the examples and embodiments describedherein are for illustrative purposes only and that various modificationsor changes in light thereof will be suggested to persons skilled in theart and are to be included within the spirit and purview of thisapplication and scope of the appended claims. Thus, the followingexamples are offered to illustrate, but not to limit the claimedinvention.

[0077] The methods, NMR probes and spectrometer systems of the presentinvention are capable of detecting less then 0.5 g/L amounts of aceticacid in wine. For the analysis of acetic acid content, the acetic acidmethyl group hydrogen nuclei and the ethyl alcohol methyl group hydrogennuclei are examined, which differ in chemical shift by approximately 1ppm. This method for acetic acid quantitation does not violate the winebottle, is harmless to the bottle contents, and can be easily extendedto the exploration of other vital ingredients and or contaminants inintact wine bottles and other sealed consumable containers.

Examples 1 Determination of Acetic Acid Levels in Wine Samples

[0078] Standards Preparation

[0079] The titration experiments were performed on full bottle aceticacid standards prepared from mixtures of de-ionized water, 200 proofethyl alcohol obtained from Gold Shield Chemical Co. (Hayward, Calif.),and 99.7% glacial acetic acid purchased from EM Science (Gibbstown,N.J.). The control samples were generated by filling, or “charging”empty wine bottles with 750 mL of 12.5% (v/v) ethyl alcohol in waterhaving a selected concentration of acetic acid (ranging between about0.5 g/L and about 3.2 g/L). Sodium chloride (Fisher Scientific, Hampton,N.H.) was dissolved in 750 mL water and used as a calibration standardfor both shimming the magnetic field for nuclei with low gyromagenticratio γ and for determining the Larmor frequency of the comparativelyless sensitive ¹³C nucleus in full bottle wine samples. The tested wineswere either purchased from local markets or obtained as gifts from theUC Davis Department of Viticulture and Enology.

[0080] Experimental Set-up

[0081] The NMR experiments on sealed wine bottles were performed at 2.01T magnetic fields corresponding to a ¹H Larmor frequency of 85.78 MHzrespectively. A high field NMR spectrometer (Varian Inc. Inova 400, PaloAlto, Calif.) employing a 9.1 T magnetic field (corresponding to a ¹HLarmor frequency of 399.76 MHz. The ) was used to confirm the aceticacid concentrations measured on the low field instrument, using 500 μLaliquots of the samples.

[0082] The single resonance NMR spectrometer delivers rf pulses to ahigh power amplifier connected to the NMR probe head mounted inside ofan Oxford Instruments (Palo Alto, Calif.) 310 mm room temperature boresuperconducting solenoid imaging magnet. The full intact wine bottle ishoused inside of the NMR probe head as shown in FIG. 3A. The rf coil isproximal to the neck of the wine bottle between the base of the cork andthe main body of the wine bottle.

[0083] Careful adjustment of the cryogenic and room temperature magneticfield shims was performed to establish a homogeneous field over the winebottle (as indicated by a ¹H line width of ≦4 Hz). Although there isless sample in this region of the bottle in comparison to the bottlebody and base, it is much easier to establish a homogeneous staticmagnetic field over the small sample region and ultimately producenarrow, highly resolved NMR lines. Current for the room temperatureshims was provided by a General Electric Omega series NMR spectrometermagnetic field shim power supply, modified to output −5 V to +5 V DC oneach channel. The supply was controlled by a potentiometer bank obtainedfrom a Varian EM 390 (90 MHz) continuous wave NMR spectrometer.

[0084] After termination of the rf pulse, the sample emits a low μV−mVrf signal that is mixed to audio frequencies and digitized by the NMRspectrometer. Fourier transformation of this signal yields the standardNMR spectrum. Substantial improvements in dynamic range can be made byselectively exciting and measuring just the methyl group region of the¹H NMR spectrum between 1 and 2.5 ppm. Operation in this way removes themassive background signal from water at 4.8 ppm.

[0085] Several different nuclei including deuterium (²H), oxygen (¹⁷O),carbon (¹³C), and hydrogen (¹H) were considered as possible candidatesfor determining acetic acid levels in wine. Of these possibilities, the¹H nucleus was chosen due to its superior sensitivity and the 1 ppmchemical shift difference between the spectrum of acetic acid and thespectra of water and ethyl alcohol, the two major constituents of wine.

[0086] NMR Data Collection: Control Data

[0087] The presence and/or quantity of acetic acid in various wine-basedsamples was determined by ¹H NMR spectroscopy at 9.1 T as a control.FIG. 5A provides a portion of an NMR spectrum generated for a 500 μLsample of the 1997 vintage UC Davis Experimental Vineyard CabernetSauvignon. The intense peak at 4.8 ppm is due to water, while thequartet and triplet centered at 3.6 ppm and 1.1 ppm represent themethylene and methyl groups in ethyl alcohol, respectively. The ¹H NMRspectrum in FIG. 5B (also obtained at 9.1 T) corresponds to a homemadesample of red wine vinegar. The new peak at approximately 2 ppm clearlyindicates the methyl group in acetic acid, and the lack of splittings ofthis single line is consistent with the chemical structure. The amountof acetic acid in the red wine vinegar was determined to be 2.6% (or27.6 g/L), based upon the ratio of the methyl group peak heights in FIG.5B, assuming that the ethyl alcohol was 12.5% of the full bottle volumeprior to acetification.

[0088] NMR Data Collection: Experimental Data

[0089] Exemplary data collected by the methods and probes of the presentinvention is shown in FIGS. 6A and 6B. The ¹H NMR spectrum in FIG. 6Awas obtained for a full bottle of the UC Davis Cabernet Sauvignon, withselective excitation of the methyl group frequencies between ±3 ppm. Thetriplet-splitting of the methyl resonance depicted in FIG. 6A(corresponding to the triplet shown at 1.1 ppm in the 500 μL sample ofFIG. 5A) is due to scalar coupling with the protons in the methylenegroup in the ethyl alcohol molecule. The full bottle ¹H NMR spectrumshown in FIG. 6B, corresponds to a 750 mL mixture of water, 12.5% ethylalcohol, and 0.5% acetic acid. The singlet peak centered at 2.1 ppm(present in the FIG. 6B vinegar sample but not the FIG. 6A wine sample)clearly indicates the presence of acetic acid (as expected) fromcomparison to the spectrum obtained for the small volume shown in FIG.5B. The NMR spectrum in FIG. 6B corresponds to an acetic acidconcentration of 5.3 g/L, nearly 3.8 times the accepted 1.4 g/L aceticacid spoilage limit for wine.

[0090] Titration Data

[0091] The titration data shown in FIG. 7 provides a comparison of theprepared acetic acid concentrations versus NMR measurements of aceticacid concentration in the prepared samples, as determined from the ratioof the integrated area of the acetic acid peak at 2.1 ppm to theintegrated area of the ethyl alcohol triplet at 1.1 ppm given the 12.5%(v/v) ethyl alcohol concentration. The open circles correspond to theaverage of nine measurements of the acetic acid concentration from fullbottle NMR spectra at 2.01 T, while the open triangles represent onemeasurement of the acetic acid concentration in a 500 μL sample at 9.1T. The dashed line of unit slope is included in FIG. 7 indicate thecorrelation between prepared and experimentally-determinedconcentrations of acetic acid. Both the low field “full bottle”measurements and the high field “small sample” measurements of aceticacid agree with prepared concentrations, although there is some spreadin the data. In the case of the high field small sample results, theuncertainty between the prepared and measured concentrations is mostlikely due to a liquid volume measurement error in the samplepreparation, as the extremely narrow ¹H NMR line widths as shown in FIG.5 permit reasonably accurate peak intensity calculation by integration.The increased line widths in the full bottle experiment shown in FIG. 6introduce more error into the measurement of acetic acid concentrationas shown by the error bars in FIG. 7, due to the increased difficulty inassigning starting and ending points for peak integration. Consequentlyerrors in both liquid volume measurements during sample preparation andpeak intensity determination introduce slightly deviations from exactagreement with the dashed line in FIG. 7. Improved magnetic field shimsyielding narrower lines will substantially increase the accuracy of theacetic acid concentration as measured. However, despite this smalldisparity, the full bottle method is capable of evaluating the amount ofwine acetification down to at least 0.5 g/L, more than half the acceptedspoilage limit of 1.4 g/L.

[0092] Further calculations

[0093] Even though wine is an extremely complex mixture of diversechemical constituents, a wine sample produces a relatively simple ¹H NMRspectrum. In the absence of spoilage, the ¹H NMR spectrum of a sample ofwine (as obtained following a single pulse excitation using the sequenceprovided in FIG. 8A) has a singlet resonance positioned at 4.8 ppm(corresponding to water), as well as an ethanol-derived quartetresonance and triplet resonance centered at 3.6 ppm and 1.1 ppm,respectively. The presence of low levels of acetic acid due to winespoilage is indicated by another singlet resonance, positioned at 2.1ppm. Taking the ratio of the integrated intensities of the ethanoltriplet to the water peak, and the acetic acid peak to the ethanoltriplet allows the percentage of ethanol by volume and the concentrationof acetic acid in wine to be quantified as: $\begin{matrix}{{{EtOH}\quad \% \quad \left( {v\text{/}v} \right)} = \frac{f_{EtOH} \times 10^{3}}{{\left( {8.5 + {8.2\quad f_{HOAc}}} \right)f_{EtOH}} + 4.6}} \\{and} \\{{\lbrack{HOAc}\rbrack \quad \left( {g\text{/}L} \right)} = \frac{f_{HOAc}f_{EtOH} \times 10^{4}}{{\left( {8.3 + {8.0\quad f_{HOAc}}} \right)f_{EtOH}} + 4.5}}\end{matrix}$

[0094] where the molecular weights and densities of water, ethanol andacetic acid have been used to calculate the values in the denominator ofthe equations. The measurement of f_(EtOH) is derived from datacollected by a one pulse experiment as depicted in FIG. 8A. However, asimilar estimate of f_(HOAc) is complicated by the strong water andethanol signals (e.g., 99% of the spectral intensity). Since the methylgroup resonance for ethanol and acetic acid are centered at 1.1 ppm and2.1 ppm respectively, and that the water resonance is shifted 2.7 ppmdownfield from the acetic acid peak (e.g., a 232 Hz downfield shift at2.01 T), the pulse sequence provided in FIG. 8B can optionally be usedfor data generation. The combination of selective excitation, delayedacquisition and block averaging can be used reliably and reproducibly tomeasure f_(HOAc) (see Weekley et al. (March 2003) “Using NMR to studyfull intact wine bottles” J. Magn. Reson. 161:91-98). The 3 ms soft rfpulse “tips” the water magnetization by less than 5 degrees, and whencombined with a 200 Hz audio filter bandwidth, the signal intensity ofthe water peak is attenuated about an order of magnitude. The delayedacquisition combined with the long spin-spin relaxation times for themethyl protons in ethanol and acetic acid reduces the short-lived freeinduction decay (fid) components that lead to broad spectral lines, thusyielding the desired narrow resonances (e.g., line widths of approx. 4Hz).

[0095] The methyl group region of the ¹H NMR spectrum for a full bottleof 1997 vintage UC Davis Cabernet Sauvignon is shown in FIG. 6A, whilethe comparable data for a full bottle having 12.5% (w/v) ethanoldissolved into water, with 0.5% (v/v) added acetic acid is shown in FIG.6B. The triplets in these spectra correspond to the ethanol methylgroup, based upon both the 1.1 ppm chemical shift and the splittingpattern (due to scalar coupling with the two equivalent methylene ¹Hnuclei in the ethanol structure). The single peak at 2.1 ppm in thespectrum shown in FIG. 6B corresponds to acetic acid. Using the formulasprovided above, f_(EtOH) is determined to be 6.4×10⁻². The ratio of theintegrated intensity of the acetic acid peak to the ethanol triplet inFIG. 6B gives f_(HOAc) as 4.5×10⁻², which can be used to calculate thatthe concentration of acetic acid [HOAc] in the sample is 5.7 g/L, ascompared to the solution as prepared (5.3 g/L of acetic acid in the 0.5%(v/v) standard solution). The 0.4 g/L difference between thesemeasurements is probably due to error in the standard preparation.

Example 2 Determining Acetic Acid Spoilage in Unopened Bottles of Wine

[0096] In most practical applications, there is no prior knowledge ofthe ration f_(EtOH), because wines of different vintages, sources, typesand quality can differ in ethanol concentration between about 7% to 24%(v/v). In these situations, the pulse sequence as provided in FIG. 8A isfirst used to measure the entire ¹H NMR spectrum, followed byapplication of the pulse sequence of FIG. 8B to selectively excite anddetect the methyl group region. In this manner, both f_(EtOH) andf_(HOAc) can be measured peak integrals and used to calculate thepercentage of ethanol and concentration of acetic acid. As noted above,data collection is typically performed via block averaging (e.g., asblock averages of n=10 groups of 100 scans. The sets of free inductionsignals are Fourier transformed, overlapped by shifting the frequency,and added offline. This procedure eliminates the effect of the long timedrift in the static magnetic field on the collected data, therebyproducing highly resolved ¹H NMR spectra for the methyl group region inwine, which can be used to accurately measure f_(HOAc).

[0097] The accuracy and sensitivity of this approach has been tested infull bottles by comparing the NMR-derived concentrations to actualprepared concentrations. The one-to-one agreement between the differentconcentration measurements with the less than 0.1 g/L acetic acidsensitivity of the full bottle NMR approach prompts further analysis.The NMR-derived percentages of ethanol (FIG. 10A) and acetic acidconcentrations (FIG. 10B) in a vertical series of sealed full bottles ofthe UC Davis Cabernet Sauvignon bottled between 1950 and 1977 werecompared. As expected, the amount of ethanol in this series does notcorrelate well with the year, and varies between 10-20%. Interestingly,the two most recent vintages display concentrations of ethanol veryclose to the industry standard for most wines (12.5% v/v). A similarlack of correlation is observed (FIG. 10B) for the full bottle aceticacid concentrations for these same wines. Although he oldest winedisplays the largest degree of acetic acid spoilage (6.3 g/L), and theyoungest wine has no measurable acetic acid contamination, the acidconcentration in the other vintages caries between 0.4 g/L and 2.0 g/L.It is therefore incorrect to assume that older wines will automaticallyhave a higher concentration of acetic acid as compared to younger wines.The integrity of the cork (and hence the quality of the bottle sealagainst oxygen leakage with time) is of paramount importance to aceticacid contamination.

[0098] It should be emphasized that the apparatus is capable ofinvestigating a variety of common bottle shapes and sizes, as well asother sealed consumables containers. All of these factors including theeffects of lead or metallic seals can be compensated for by carefullyadjusting the home built room temperature magnetic field shims.Additionally, the lead or metallic seals do not measurably interferewith the probe tuning or the homogeneity and intensity of the rf fieldacross the wine bottle. Although the titration data shown in FIG. 7 onlydocuments results down to 0.5 g/L acetic acid, levels down to 0.1 g/Lhave been measured with the probes and systems of the present invention.It is anticipated that NMR solvent suppression techniques and/or a dualcoil NMR probe head will extend the sensitivity by one or more orders ofmagnitude.

Example 3 ¹³C NMR Spectroscopy of Bottle Samples

[0099] As noted herein, the present invention for the NMR analysis ofsealed consumables containers are not limited to methods and devicesinvolving performing ¹H NMR spectroscopy. In an effort to increase thesensitivity of measurements of dilute components (like flavenoids andaldehydes), as well as to extend the full bottle technique to nucleiother than ¹H, an additional probe embodiment was constructed (see FIGS.4A and 4B). Instead of examining the approximately 25 cm³ sample volumein the neck of the wine bottle, the probe can be used to analyze themuch larger (˜1L) volume in the body of the wine bottle. Although themagnetic field homogeneity is worse across a larger sample volume,examination of nuclei having a larger chemical shift dispersion than ¹Hwill be less sensitive to the increased line width.

[0100] In one embodiment of the methods of the present invention, sealedconsumables containers are examined using ¹³C NMR spectroscopy. The muchwider chemical shift range and lower Larmor frequency of ¹³C as comparedto ¹H (21.56 MHz versus 85.78 MHz at 2.01 T, respectively) reduced thenecessity for narrow line width for analysis. As such, it becomesfeasible to center the rf detection coil on the main body of the winebottle, thereby improving sensitivity (due to greater volume of nuclei)without sacrificing the rf coil filling factor.

[0101] The formation of spin echoes for low γ nuclei is possible usingthe probes of the present invention (see, for example, FIGS. 4A and 4B),despite the observation that the geometry of the four turn splitsolenoid rf coil is not optimized for homogeneity. In the special caseof ¹³C NMR spectroscopy, in which the spin-lattice and spin-spinrelaxation times tend to be long, multiple π pulse sequences (asdepicted in FIG. 8C) can be employed to refocus the magnetization andincrease the signal to noise ratio (S/N) for a fixed number of scans byadding (offline) the free induction signal following the 100 μs π/2pulse to the echo signals appearing at 102 ms intervals. In this manner,fully ¹H-coupled ¹³C NMR spectra corresponding to 100-1000 scans can beobtained for full bottle samples in a reasonable period of time

[0102]FIGS. 9A and 9B depict ¹³C spectra on full bottles of either the1997 UC Davis Cabernet Sauvignon (9A) or red wine vinegar (9B), usingthe pulse sequence provided in FIG. 8C with n=7. The triplet and quartetcentered at 57 ppm and 18 ppm arise from the methylene and methylcarbons of ethanol, respectively. The line splitting of about 140 Hz inboth of these peaks, as well as their splitting patterns, are consistentwith scalar coupling to directly bonded ¹H nuclei. In the vinegarsample, additional ¹³C peaks are seen at 18 ppm and 21 ppm, due to thecarbonyl and methyl groups of the acetic acid. The inverted triangles inFIG. 10B label the acetic acid methyl group quartet. The near-equalintegrated intensity of the nested quartets suggests that the amount ofethanol and acetic acid in the sample of red wine vinegar are nearlyequal, a result consistent with the literature (Jakish (1985) ModernWinemaking Cornell University Press, Ithaca N.Y.).

[0103] It is clear from the spectra that the full bottle ¹³C NMR methodis feasible for the exploration of additional wine components, such astannins, flavenoids, phenols, aldehydes and amino acids. In principle,continues signal averaging will reveal these peaks in the ¹³C spectrum,although the spectra will be very complicated in the absence ofdecoupling from the ¹H nuclei. Optionally, an additional ¹H channel isincorporated into the probes of the present invention, thereby providingincreased resolution and sensitivity (and potentially, nuclearOverhauser effects) through the use of ¹H decoupling. Furthermore, probeembodiments for detection of additional isotopes, such as ²⁰⁷Pb, ¹⁹⁹Hg,⁴⁵Sc, ³⁹K, ²⁷Al, ²³Na and the like are also contemplated. Although theabundance of these isotopes is typically below the detection limit forstandard (i.e., microliter volume) NMR spectroscopy, the increasedvolumes employed in the full bottle spectroscopic methods and probesamplifies the number of spins by a factor of 10⁴, thus making the studyof trace elements in native wine samples accessible for the first time.Moreover, the methods and devices of the present invention can be usedto analyze the quality and nature of the wine bottle itself (e.g., by acombination of ²⁹Si and ²³Na NMR spectroscopy), while the cork (eithernatural or synthetic) could be studied, e.g., using ¹³C solid state NMRtechniques.

[0104] The discussion above is generally applicable to the aspects andembodiments of the present invention. Moreover, modifications can bemade to the methods, apparatus, and systems described herein withoutdeparting from the spirit and scope of the invention as claimed, and theinvention can be put to a number of different uses including thefollowing:

[0105] The use of an NMR probe configured to accept a sealed consumablescontainer or an NMR system as set for the herein, for performing any ofthe methods and assays set forth herein.

[0106] The use of an NMR probe or system as described herein forperforming noninvasive analysis of a corked wine bottle or any othersealed consumables container, e.g., for analysis of one or morecontaminants, as set forth herein.

[0107] A kit comprising one or more standard solutions of contaminant(e.g., acetic acid titration samples) in a sealed consumables container,for use in the methods, devices or systems of the present invention.Optionally, the kit further comprises an instruction manual forperforming the methods of the present invention.

[0108] While the foregoing invention has been described in some detailfor purposes of clarity and understanding, it will be clear to oneskilled in the art from a reading of this disclosure that variouschanges in form and detail can be made without departing from the truescope of the invention. For example, all the techniques and apparatusdescribed above can be used in various combinations. All publications,patents, patent applications, and/or other documents cited in thisapplication are incorporated by reference in their entirety for allpurposes to the same extent as if each individual publication, patent,patent application, and/or other document were individually indicated tobe incorporated by reference for all purposes.

What is claimed is:
 1. A method of analyzing one or more contents of asealed consumables container, the method comprising: providing an NMRspectrometer and an NMR probe configured to accept a portion of thesealed consumables container; positioning the portion of the containerwithin a data collection region of the NMR probe; establishing ahomogeneous static magnetic field across the data collection region;collecting an NMR spectrum; and analyzing one or more peaks in the NMRspectrum, thereby analyzing one or more contents of the sealedconsumables container.
 2. The method of claim 1, wherein the sealedconsumables container comprises a bottle of wine.
 3. The method of claim1, wherein the sealed consumables container comprises a container ofnonalcoholic beverage, alcoholic beverage, beer, vinegar or olive oil.4. The method of claim 1, wherein the sealed consumables container holdscomprises a food or beverage container, wherein the contained food orbeverage comprises components having sharply defined NMR peaks.
 5. Themethod of claim 1, wherein analyzing the container comprises determininga presence or a concentration of a selected component of the contents.6. The method of claim 5, wherein the selected component comprisesacetic acid.
 7. The method of claim 5, wherein the selected componentcomprises one or more aldehydes.
 8. The method of claim 5, wherein theselected component comprises one or more flavenoids.
 9. The method ofclaim 1, wherein positioning the portion of the container comprisesplacing a neck of the container within the data collection region of theNMR probe.
 10. The method of claim 1, wherein positioning the portion ofthe container comprises placing a body of the container within the datacollection region of the NMR probe.
 11. The method of claim 1, whereinthe NMR spectrometer further comprises one or more shim coils, andwherein establishing the homogeneous static magnetic field across thedata collection region comprises adjusting the one or more shim coils.12. The method of claim 1, wherein analyzing the one or more peakscomprises performing integration on the one or more peaks.
 13. An NMRprobe configured to position a portion of a sealed consumables containerwithin an NMR spectrometer, the probe comprising: a body structurehaving a cavity disposed at a first end of the body structure, saidcavity being adapted for receiving a portion of the sealed container; afirst rf coil positioned proximal to the cavity and the portion of thesealed container; and a tuning capacitor coupled at a first position tothe rf coil and coupled at a second position to a length of coaxialcable configured for connection to the NMR spectrometer.
 14. The NMRprobe of claim 13, wherein the probe comprises a ¹H probe.
 15. The NMRprobe of claim 13, wherein the probe comprises a ²H probe.
 16. The NMRprobe of claim 13, wherein the probe comprises a ¹³C probe.
 17. The NMRprobe of claim 13, wherein the probe comprises a ¹⁷O probe.
 18. The NMRprobe of claim 13, wherein the first rf coil comprises 12 gauge copperwire wound as an eight turn 1 cm diameter split solenoid coil.
 19. TheNMR probe of claim 18, wherein the first rf coil comprises a first coilportion having 4 turns of copper wire positioned at a first side of thecavity, and a second coil portion having 4 turns of copper wire andcoupled to the first coil portion by a copper connecting wire, whereinthe second coil portion is positioned on an opposite side of the cavityand along a same axis as the first coil portion, whereby the two coilportions encompass the portion of the sealed container.
 20. The NMRprobe of claim 18, wherein the first rf coil comprises a birdcage-stylecoil.
 21. The NMR probe of claim 13, wherein the first rf coil comprisesa coil used for both transmitting and receiving rf pulses.
 22. The NMRprobe of claim 13, further comprising a second rf coil positioned distalto the first rf coil.
 23. The NMR probe of claim 22, wherein the secondrf coil is configured for measurement of one or more signals from acalibration sample.
 24. The NMR probe of claim 22, wherein the second rfcoil is configured for selective excitation of a heteronucleus.
 25. TheNMR probe of claim 23, wherein the heteronucleus comprises ¹³C, ¹⁷O, ²H,²³Na, ²⁷Al, ¹⁹⁹Hg, or ²⁰⁷Pb.
 26. The NMR probe of claim 13, furthercomprising components for generating one or more magnetic fieldgradients, wherein the components are coupled to the body structure. 27.The NMR probe of claim 26, wherein the components for generating one ormore magnetic field gradients comprise imaging components.
 28. The NMRprobe of claim 13, wherein the portion of the sealed consumablescontainer comprises a neck of the container.
 29. The NMR probe of claim13, wherein the portion of the sealed consumables container comprises abody of the container.
 30. The NMR probe of claim 13, further comprisinga tuning capacitor coupled to the first rf coil, wherein the tuningcapacitor comprises one or more non-magnetic 0-10 picofarad high powerrf capacitors.
 31. The NMR probe of claim 13, further comprising acalibration fluid sample tube positioned within the cavity of the bodystructure and adjacent to the portion of the sealed consumablescontainer when the container is inserted in the cavity.
 32. The NMRprobe of claim 13, further comprising a fluid jacket for modulating atemperature of the probe.
 33. A system for analyzing contents of asealed consumables container, comprising: the NMR probe of claim 13; anNMR spectrometer comprising a body structure, a magnet housed within thebody structure, a bore proximal to the magnet and configured to receivethe NMR probe, thereby positioning a portion of the container within amagnetic field generated by the magnet, and an amplifier configured forcoupling to a first position on the NMR probe; and a receiver systemconfigured for electronic communication with the NMR probe, the receiversystem comprising a preamplifier configured for coupling to a secondposition on the NMR probe and a detector in communication with thepreamplifier.
 34. The system of claim 33, wherein the NMR probecomprises a single resonance probe selected from the group consisting ofa ¹H probe, a ²H probe, a ¹³C probe, an ¹⁷O probe, a ²³Na probe, a ²⁷Alprobe, a ¹⁹⁹Hg probe, and a ²⁰⁷Pb probe.
 35. The system of claim 33,wherein the first rf coil of the NMR probe comprises a coil used forboth transmitting and receiving rf pulses.
 36. The system of claim 33,wherein the NMR probe further comprises a second rf coil.
 37. The NMRprobe of claim 36, wherein the second rf coil is configured formeasurement of one or more signals from a calibration sample.
 38. Thesystem of claim 33, wherein the portion of the sealed consumablescontainer comprises a neck of the container.
 39. The system of claim 33,wherein the portion of the sealed consumables container comprises a bodyof the container.
 40. The system of claim 33, wherein the magnetcomprises a room temperature superconducting magnet.
 41. The system ofclaim 33, wherein the magnetic field comprises a 2.01 T magnetic field.42. The system of claim 33, wherein the receiver comprises a passive rfduplexer and signal mixing and digitization electronics.
 43. The systemof claim 33, further comprising an NMR pulse programmer.